Cell Preservation Services, Inc. (CPSI) is a biotechnology company dedicated to the development of novel technologies in the area of low temperature biology and medicine. CPSI's core molecular strategy is focused on understanding and manipulating the cell survival and cell death pathways that appear to be cell-specific and are activated as a consequence of low temperature exposure. This paradigm shift has led to improved processes for the hypothermic storage and cryopreservation of a variety of human cells and tissues. Recently, through preliminary studies, we have determined that it is not adequate to merely address stress response issues during preservation through solution design, as has been the focus in the past, but in fact a more comprehensive and integrated strategy involving pre- and post-preservation cell conditioning strategies is necessary. CPSI applied this novel strategic approach and has identified promising new methodologies that appear to improve corneal cell and tissue quality beyond that of typical protocols which rely only on the use of preservation media, such as Optisol-GS, to maintain tissue integrity and function. Feasibility data gathered on human corneal endothelial cells (HCEC), bovine corneal endothelial cells (BCEC) and other relevant human cell strains suggest that the development of improved cornea processing and preservation protocols is highly feasible and successful development of an integrated process should not only increase the quality and utility of corneal tissue, but increase the donor pool as well. Investigations conducted under this research project will utilize cDNA microarrays, proteomic profiling, and viability and functional analysis to define the stress response pathways activated in HCEC following low temperature exposure common to pre-transplant processing. It is the intent of these studies to provide a better understanding of the unique responses of HCEC and whole cornea to low temperature exposure and utilize these data to develop new procedures for corneal tissue processing. We have established collaborations and subcontracts with researchers from several organizations including Harvard Medical School, the Schepens Eye Research Institute, the Moran Eye Center and NDRI to facilitate access to various outside resources (including expertise, cell culture systems, IP, etc.) to evaluate new processes/protocols that are developed under this research project as well as to help ensure the success of this research program. It is our belief that the proposed investigations will lead to new methodologies for the processing of corneal tissue. These developments should have an impact not only on the quality and availability of harvested corneas, but also enable expanded utilization of corneal-derived cells in areas such as in vitro toxicological testing and HCEC transplant. Corneal opacities are leading causes of blindness. One method to resolve this impediment to clear vision is to transplant donor corneas to the patient. To do so typically means that donor corneas must be preserved in a state of suspended animation in the cold prior to transplant. The methodologies developed in the past using Optisol- GS are dated and not based on modern, molecular medicine. CPSI proposes to use a molecular strategy to develop improved protocols for the long term storage of human corneas - a set of integrated processes that will both increase the available cornea donor pool as well as the quality of the tissue for the cornea transplant market.